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Effect of stirring speed on the production of phenolic secondary metabolites and growth of Buddleja cordata cells cultured in mechanically agitated bioreactor

  • Alicia Monserrat Vazquez-Marquez
  • Carmen Zepeda-Gómez
  • Cristina Burrola-Aguilar
  • Antonio Bernabé-Antonio
  • Aurelio Nieto-Trujillo
  • Francisco Cruz-Sosa
  • Mario Rodríguez-Monroy
  • María Elena Estrada-ZúñigaEmail author
Original Article
  • 19 Downloads

Abstract

Shake-flask in vitro culture of Buddleja cordata cells produces large amounts of biomass and synthetizes verbascoside (VB), linarin and hydroxycinnamic acids, bioactive phenolic secondary metabolites (PSMs). In this work, we determined the effect of stirring speed on the growth of and production of PSMs [total phenolic, phenylethanoid glycoside and flavonoid contents (PeC, PeGC and FC, respectively)] by B. cordata cells cultured in two bioreactors. Two different stirring speeds (120 and 400 rpm) were tested in two stirred-tank bioreactors: a 2 L bioreactor equipped with a ring diffuser (B2RD) and a 3 L bioreactor with a sintered diffuser (B3SD). Growth kinetics of B. cordata cells were measured in the bioreactors and shake-flask systems. The stirring speed and type of bioreactor affected phases, parameters of growth and production of PSMs. The highest production of biomass (13.62 g L−1) and PSMs [PeC of 64.63 mg gallic acid equivalents g−1 (mg GAE g−1); PeGC of 119.24 mg VB equivalents g−1 (mg VBE g−1); and FC of 5.02 mg quercetin equivalents g−1 (mg QE g−1)] occurred in B2RD at 400 rpm. These values were similar to the found in shake-flasks system. This work establishes the basis for bioprocess advances of B. cordata focused on the development of a sustainable strategy for the management of natural resources and as a source of bioactive PSMs on a large scale.

Key message

Buddleja cordata cells cultured in a mechanically agitated bioreactor possess an outstanding biosynthetic potential that represents a suitable biotechnological alternative for the production of bioactive phenolic secondary metabolites.

Keywords

Bioreactor Buddleja cordata Phenolic secondary metabolites Diffuser Stirring speed Verbascoside 

Abbreviations

µ

Specific growth rate

ANOVA

Analysis of variance

B2RD

2 L Bioreactor equipped with a ring diffuser

B3SD

3 L Bioreactor equipped with a sintered diffuser

CSCBc

Cell suspension culture of B. cordata

CV

Cellular viability

DO

Dissolved oxygen

DW

Dry weight

F

Flavonoid (s)

FC

Total flavonoid content

GI

Growth index

GAE

Gallic acid equivalent

MB

Maximum biomass

MSDK

Murashige and Skoog-modified culture medium

Pe

Phenolic compound (s)

PeC

Total phenolic content

PeG

Phenylethanoid glycoside (s)

PeGC

Total phenylethanoid glycoside content

PSM

Phenolic secondary metabolite (s)

QE

Quercetin equivalent

qpF

Specific production rate of total flavonoids

qpPe

Specific production rate of total phenolics

qpPeG

Specific production rate of total phenylethanoid glycosides

RA

Rosmarinic acid

SM

Secondary metabolite (s)

td

Doubling time

TSC

Total sugar content

VB

Verbascoside

VBE

Verbascoside equivalent

Yx/s

Yield of biomass from substrate

Notes

Acknowledgements

The authors thank the Universidad Autónoma del Estado de México (UAEM) for financing this thesis project through the Programa de Investigación Científica, Innovación y Desarrollo UAEM 2014 (Project No. 3742/2014/CIB: Desarrollando Avances Biotecnológicos Sobre la Producción de Verbascósido por Cultivos de Células de Buddleja cordata) and to the Consejo Nacional de Ciencia y Tecnología (CONACyT) through grant number 620491 for the Master’s studies of AMV-M at the Posgrado en Ciencias Agropecuarias y Recursos Naturales from UAEM.

Authors contributions

AMV-M, as a Master student and for her thesis project, participated in all the experimental work, analysis and interpretation of data, and writing of this manuscript. CZ-G supervised the establishment of the experiments and provided chemical standards. CB-A participated in the standardization of analytical procedures to quantify the secondary metabolites. AB-A participated in the analysis of the growth kinetics and their statistical analysis. AN-T participated in the experimental work on phytochemical analysis. FC-S contributed to the design and execution of experiments on shake-flask cultures; participated in the analysis and interpretation of data. MR-M contributed to the thesis project of MAV-M as an assessor; supervised the establishment of bioreactor in vitro cultures; contributed to the planning and execution of this project, which led to this publication; and contributed to the preparation and writing of this manuscript. MEE-Z contributed to the design, execution and direction of all the experiments of this project, as she was the thesis project director of AMV-M; was the responsible for the design and direction of Project No. 3742/2014/CIB, which financially supported this thesis project; and critically contributed to the preparation of the manuscript until approving the final submitted version. All authors critically reviewed the manuscript and approved the final version.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Facultad de CienciasUniversidad Autónoma del Estado de MéxicoTolucaMexico
  2. 2.Centro de Investigación en Recursos Bióticos-Facultad de CienciasUniversidad Autónoma del Estado de MéxicoTolucaMexico
  3. 3.Departamento de Madera, Celulosa y Papel, Centro Universitario de Ciencias Exactas e IngenieríasUniversidad de GuadalajaraZapopanMexico
  4. 4.Departamento de BiotecnologíaUniversidad Autónoma Metropolitana-IztapalapaCol. VicentinaMexico
  5. 5.Departamento de BiotecnologíaCentro de Desarrollo de Productos Bióticos del Instituto Politécnico Nacional, YautepecMorelosMexico

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